Long noncoding RNA and microRNA are regulatory noncoding RNAs that are implicated in Alzheimer's disease, but the role of long noncoding RNA-associated competitive endogenous RNA has not been fully elucidated. The...Long noncoding RNA and microRNA are regulatory noncoding RNAs that are implicated in Alzheimer's disease, but the role of long noncoding RNA-associated competitive endogenous RNA has not been fully elucidated. The long noncoding RNA growth arrest-specific 5(GAS5) is a member of the 5′-terminal oligopyrimidine gene family that may be involved in neurological disorders, but its role in Alzheimer's disease remains unclear. This study aimed to investigate the function of GAS5 and construct a GAS5-associated competitive endogenous RNA network comprising potential targets. RNA sequencing results showed that GAS5 was upregulated in five familial Alzheimer's disease(5×FAD) mice, APPswe/PSEN1dE9(APP/PS1) mice, Alzheimer's disease-related APPswe cells, and serum from patients with Alzheimer's disease. Functional experiments with targeted overexpression and silencing demonstrated that GAS5 played a role in cognitive dysfunction and multiple Alzheimer's disease-associated pathologies, including tau hyperphosphorylation, amyloid-beta accumulation, and neuronal apoptosis. Mechanistic studies indicated that GAS5 acted as an endogenous sponge by competing for microRNA-23b-3p(miR-23b-3p) binding to regulate its targets glycogen synthase kinase 3beta(GSK-3β) and phosphatase and tensin homologue deleted on chromosome 10(PTEN) expression in an Argonaute 2-induced RNA silencing complex(RISC)-dependent manner. GAS5 inhibited miR-23b-3p-mediated GSK-3β and PTEN cascades with a feedforward PTEN/protein kinase B(Akt)/GSK-3β linkage. Furthermore, recovery of GAS5/miR-23b-3p/GSK-3β/PTEN pathways relieved Alzheimer's disease-like symptoms in vivo, indicated by the amelioration of spatial cognition, neuronal degeneration, amyloid-beta load, and tau phosphorylation. Together, these findings suggest that GAS5 promotes Alzheimer's disease pathogenesis. This study establishes the functional convergence of the GAS5/miR-23b-3p/GSK-3β/PTEN pathway on multiple pathologies, suggesting a candidate therapeutic target in Alzheimer's disease.展开更多
[目的]探讨miR-23b-3p靶向调节TFRC对子宫内膜癌Ishikawa细胞转移活性的影响。[方法]设Ishikawa子宫内膜癌细胞组、miRNA-NC组、miR-23b-3p inhibitor组、miR-23b-3p mimics组,培养72h后,测定细胞活力、凋亡及细胞侵袭迁移水平,RT-PCR...[目的]探讨miR-23b-3p靶向调节TFRC对子宫内膜癌Ishikawa细胞转移活性的影响。[方法]设Ishikawa子宫内膜癌细胞组、miRNA-NC组、miR-23b-3p inhibitor组、miR-23b-3p mimics组,培养72h后,测定细胞活力、凋亡及细胞侵袭迁移水平,RT-PCR法、蛋白印迹法测定细胞miR-23b-3p、TFRC水平。[结果]与Ishikawa子宫内膜癌细胞组、miRNA-NC组比较,miR-23b-3p inhibitor组OD值(0.84±0.05 vs 0.83±0.06 vs 0.93±0.07,P<0.05)、存活率(88.85%±2.33%vs 86.34%±2.74%vs 95.27%±2.25%,P<0.05)、穿膜数、迁移距离(18.36±5.25 vs 19.33±6.30 vs 65.63±12.38,P<0.05)升高,凋亡率降低(6.11%±1.25%vs 6.83%±1.18%vs 1.13%±0.24%,P<0.05);miR-23b-3p mimics组的Ishikawa细胞的OD值、存活率、穿膜数、迁移距离降低,凋亡率升高(P<0.05);与miR-23b-3p inhibitor组比较,miR-23b-3p mimics组的Ishikawa细胞的OD值、存活率、穿膜数、迁移距离降低,凋亡率升高(P<0.05)。与Ishikawa子宫内膜癌细胞组、miRNA-NC组比较,miR-23b-3p inhibitor组的Ishikawa细胞的miR-23b-3p表达水平降低,TFRC mRNA水平和蛋白表达水平升高(0.95±0.15 vs 1.78±0.11,P<0.05),miR-23b-3p mimics组的Ishikawa细胞的miR-23b-3p表达水平升高,TFRC mRNA和蛋白表达水平降低(0.95±0.15 vs 0.34±0.17,P<0.05);与miR-23b-3p inhibitor组比较,miR-23b-3p mimics组的Ishikawa细胞的miR-23b-3p表达水平升高,TFRC mRNA和蛋白表达水平降低(1.78±0.11 vs 0.34±0.17,P<0.05)。[结论]miR-23b-3p靶向抑制TFRC表达进而抑制子宫内膜癌Ishikawa细胞的转移。展开更多
基金supported by the National Natural Science Foundation of China,Nos. 82173806 and U1803281Chinese Academy of Medical Sciences (CAMS) Innovation Fund for Medical Science,Nos. 2021-I2M-1-030 and 2022-I2M-2-002Non-Profit Central Research Institute Fund of Chinese Academy of Medical Sciences,No. 2022-JKCS-08 (all to RL)。
文摘Long noncoding RNA and microRNA are regulatory noncoding RNAs that are implicated in Alzheimer's disease, but the role of long noncoding RNA-associated competitive endogenous RNA has not been fully elucidated. The long noncoding RNA growth arrest-specific 5(GAS5) is a member of the 5′-terminal oligopyrimidine gene family that may be involved in neurological disorders, but its role in Alzheimer's disease remains unclear. This study aimed to investigate the function of GAS5 and construct a GAS5-associated competitive endogenous RNA network comprising potential targets. RNA sequencing results showed that GAS5 was upregulated in five familial Alzheimer's disease(5×FAD) mice, APPswe/PSEN1dE9(APP/PS1) mice, Alzheimer's disease-related APPswe cells, and serum from patients with Alzheimer's disease. Functional experiments with targeted overexpression and silencing demonstrated that GAS5 played a role in cognitive dysfunction and multiple Alzheimer's disease-associated pathologies, including tau hyperphosphorylation, amyloid-beta accumulation, and neuronal apoptosis. Mechanistic studies indicated that GAS5 acted as an endogenous sponge by competing for microRNA-23b-3p(miR-23b-3p) binding to regulate its targets glycogen synthase kinase 3beta(GSK-3β) and phosphatase and tensin homologue deleted on chromosome 10(PTEN) expression in an Argonaute 2-induced RNA silencing complex(RISC)-dependent manner. GAS5 inhibited miR-23b-3p-mediated GSK-3β and PTEN cascades with a feedforward PTEN/protein kinase B(Akt)/GSK-3β linkage. Furthermore, recovery of GAS5/miR-23b-3p/GSK-3β/PTEN pathways relieved Alzheimer's disease-like symptoms in vivo, indicated by the amelioration of spatial cognition, neuronal degeneration, amyloid-beta load, and tau phosphorylation. Together, these findings suggest that GAS5 promotes Alzheimer's disease pathogenesis. This study establishes the functional convergence of the GAS5/miR-23b-3p/GSK-3β/PTEN pathway on multiple pathologies, suggesting a candidate therapeutic target in Alzheimer's disease.
文摘[目的]探讨miR-23b-3p靶向调节TFRC对子宫内膜癌Ishikawa细胞转移活性的影响。[方法]设Ishikawa子宫内膜癌细胞组、miRNA-NC组、miR-23b-3p inhibitor组、miR-23b-3p mimics组,培养72h后,测定细胞活力、凋亡及细胞侵袭迁移水平,RT-PCR法、蛋白印迹法测定细胞miR-23b-3p、TFRC水平。[结果]与Ishikawa子宫内膜癌细胞组、miRNA-NC组比较,miR-23b-3p inhibitor组OD值(0.84±0.05 vs 0.83±0.06 vs 0.93±0.07,P<0.05)、存活率(88.85%±2.33%vs 86.34%±2.74%vs 95.27%±2.25%,P<0.05)、穿膜数、迁移距离(18.36±5.25 vs 19.33±6.30 vs 65.63±12.38,P<0.05)升高,凋亡率降低(6.11%±1.25%vs 6.83%±1.18%vs 1.13%±0.24%,P<0.05);miR-23b-3p mimics组的Ishikawa细胞的OD值、存活率、穿膜数、迁移距离降低,凋亡率升高(P<0.05);与miR-23b-3p inhibitor组比较,miR-23b-3p mimics组的Ishikawa细胞的OD值、存活率、穿膜数、迁移距离降低,凋亡率升高(P<0.05)。与Ishikawa子宫内膜癌细胞组、miRNA-NC组比较,miR-23b-3p inhibitor组的Ishikawa细胞的miR-23b-3p表达水平降低,TFRC mRNA水平和蛋白表达水平升高(0.95±0.15 vs 1.78±0.11,P<0.05),miR-23b-3p mimics组的Ishikawa细胞的miR-23b-3p表达水平升高,TFRC mRNA和蛋白表达水平降低(0.95±0.15 vs 0.34±0.17,P<0.05);与miR-23b-3p inhibitor组比较,miR-23b-3p mimics组的Ishikawa细胞的miR-23b-3p表达水平升高,TFRC mRNA和蛋白表达水平降低(1.78±0.11 vs 0.34±0.17,P<0.05)。[结论]miR-23b-3p靶向抑制TFRC表达进而抑制子宫内膜癌Ishikawa细胞的转移。
